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Pollenkitt and viscin threads: their role in

cementing pollen grains

Michael Hesse

To cite this article: Michael Hesse (1981) Pollenkitt and viscin threads: their role in cementing
pollen grains, Grana, 20:3, 145-152, DOI: 10.1080/00173138109427656

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Grana 20: 145-152, 1981

Pollenkitt and viscin threads:

their role in cementing pollen grains

hlICHAEL HESSE

Hesse, hl.: Pollenkitt and viscin threads: their role in cementing pollen grains. - Grana
20: 145-152, 1981. Uppsala 30 hlarch 1982. ISSN 0017-3134.
The principle of attachment of pollen grains to flower-visiting insects is quite different in
entomophilous angiosperms depending whether pollenkitt or viscin threads are the pollen
adhesives. Pollenkitt is to be found in all angiosperm families investigated up to now, but
viscin threads occur only in a small number. hloreover, the two pollen cementing principles
do not correspond in their respective origin, structure, sculpture, and chemical and physi-
cal characters. Even the mechanical aspect of pollen cementing differs. Thus pollenkitt and
viscin threads are analogous vehicles in pollination ecology.
Key words: Pollenkitt, viscin threads, pollen stickiness, microsporogenesis, pollination
ecology
Michael Hesse, Instiiirtfjir Botanik irnd Botanischer Garten der Universitiit Wien, Reritweg
14, A-1030 Wien, Austria.
(Mnririscript receiivtl20 October 19S0, revised version accepted20 Mnrch 1981)

Pollination means the transfer of fresh pollen from
the anthers to the female region ofgymnosperms or
angiosperms. In most gymnosperms (cf. Fzgri &
van der Pijl 1979) and all anemophilous angio-
sperms the pollen is dry, not sticky and does not
clump together, while in zoophilous (mostly en-
tomophi~ous) angiosperms the pollen transfer by
animals depends ma>inly9~ the,adherence o.f sticky,
clumping pollen graini-!@’ !hf?.nower‘viiito;. Such
pollen attachment is cciUse2- by pollen aggregation
or “pollen cementing” realized by two principally
different modes: either by the sticky, viscous ‘‘pol-
lenkitt” or by the non-sticky, non-viscous “viscin
threads”. Pollenkitt is found in all angiosperm
families investigated up to now (Hesse 1980 ( 1 ) . In
contrast, viscin threads occur only in a few, not
closely related angiosperm the Onag-
raceae, Ericaceae and Caesalpiniaceae (Skvarla e t
al. 1978, Cruden & Jensen 1979, Graham et al.
1980), “False viscin threads” which do not contain
sporopollenin occur in Strelitzicr, and some orchids
(Skvarla et al. 1978, Vijayaraghavan
~~ - & Shukla
1980). The present paper intends to review our
knowledge o n pollen aggregation and to show that
apart from their more or less equal ecological func-
tion these two principles of pollen cementing are
fundamentally different.
MATERIAL A N D hlETHODS
Pollen grains collected from anthers at various stages of
development, and also from flower visiting insects were
fixed in 6%-Sorensen-buffered glutaraldehyde, postfixed
in 2 %-OsO,, and dehydrated in ethanol after Sitte (1962)
to avoid extreme loss of lipids (see also Dunbar 1981).
After this the material was in general not acetolysed (thus
the tapetum-borne substances, especially the pollenkitt,
remain on the pollen surface), although acetolysed materi-
al was used for studying the viscin threads on the pollen
surfaces. For light microscopy thin, hand-made sections
were stained with Sudan Ill; for electron microscopy the
’ material was either embedded in Spurr’s (1969) “low vis-
cosity resin”, poststained with alcoholic uranyl acetate,
and investigated in a Zeiss Ehf-9 S-2, or critical-point-
dried, sputter-coated with gold, and investigated with a
SEhi ISI-60.
R E S U L T S AND DISCUSSION
Pollenkitt can be defined as a complex mixture of
lipid, viscous substances (including carotenoids,

10-825895 Grana 20
146 M . Hessc
Fig. I . (A) After fixation with ethanol the pollenkitt does
not cover the pollen surface of the entornophilous Bryoriio
dioica as a thin layer, but rather as strands. SEM (bar in-
dicates 10 pm). (B) In some amphiphilous angiosperms,
like Gnlnrdzrts riivolis, the pollen is fairly dry and not
sticky, as the heterogeneous and foamy pollenkitt fills up
the exine cavities (arrows), while the tectum and tectal
perforations lack pollenkitt. TEhl X31000.
but also hitherto unknown substances), produced
by the plastids of the anther tapetum and deposited
on the pollen surface and/or in the exine cavities of
pollen grains in entomophilous and anemophilous
angiosperms (strikingly pollenkitt is lacking in gym-
nosperms, even those which are entomophilous,
Hesse 1980~).Because of its ubiquity in angio-
sperms the pollenkitt (this term is often used by
German speaking authors) or the more or less
homologous tryphine (a term used often by English
speaking authors) ‘plays an important role in the
pollination ecology of entomophilous species. It
facilitates not only the cohesion of single pollen
grains after dehiscence, but also the attachment of
the pollen grains to flower-visiting insects and
moreover, acts as a store for recognition substances
in the pollination process (Dickinson & Lewis 1973)
or even as a protection from pollen dehydration.
Allergenic substances may be located in these pol-
len surface coatings. The development of pollenkitt
Grana 20
is divided into three steps (Dickinson 1973, Hesse
1978 n ) : (1) Shortly after meiosis, during and after
the tetrad stage the plastids of the tapetum pro-
duce lipid osmiophilic globuli. (2) These globuli fuse
in the tapetal cytoplasm after leaving the plastids
during and after the degeneration of the tapetal
cells. (3) Shortly before the opening of the pollen
sac takes place the fused lipid substances are de-
posited on the exine surface (e.g. the tectum) or in
the exine cavities.
The appearance of the pollenkitt sometimes de-
pends on the preparation method. In dry pollen
grains from herbarium specimens the pollenkitt ap-
pears as a very thin layer or as small lumps on the
pollen surface. After alcohol fixation the pollenkitt
sometimes has a thread-like appearance (Fig. 1 A).
This artifact must not be interpreted as “viscin
threads”, since these “pollenkitt ropes” never
coalesce with the exine, their size is very variable
lacking a constant pattern, and above all these
ropes can be easily dissolved in acetone or by
aceto!ysis. Aftcr aldchyde fixation the pollenkitt
generally appears well preserved throughout (Hesse
1 9 7 9 ~ ) .I t is a more or less viscous fluid, often
strictly homogeneous and highly electron dense
(e.g. in Horiicirrielis or in Alisrizn), but sometimes
heterogeneous. Within a dense lump of pollenkitt
either thin lipid lamellae of different electron densi-
ty, as in Plrriitngo or Cnrex, or some electron trans-
parent droplets, as in Eiiplrorbin or Arternisia, or
even small empty hollows, as in Ctrstarieo, are to be
found (Hesse 1978 b, 1979 0 , 1980 n , d). Since pol-
lenkitt in most cases is a highly viscous fluid (crys-
talloid, but nevertheless “oily inclusions”, as in
Gnlnntkia, are very rare), there is no ornamenta-
tion at all, but sometimes its surface looks like a
“crater landscape” probably due to dissolving of
some components during the preparation.
The distribution of the pollenkitt on the pollen
surface mainly influences the resulting stickiness
(or dryness) of pollen, In entomophilous angio-
sperms the mostly electron dense, opaque, highly
viscous pollenkitt makes the pollen grains sticky,
because it is mainly located on the surface of the
exine. In reticulate exines, as in Hrirririrriclis vir-
gininm (Fig. 2 A) pollenkitt covers large parts
of the reticulum as an electron dense, strictly
homogeneous crust filling up the lumina. In tectate
pollen grains on the other hand, like those ofSngit-
forin strgiffifolio, the pollenkitt covers large parts
of the tectum as a thick, very electron dense,
Fig. 2. (A) After degeneration of the tapetal cells a great
number of black droplets (pollenkitt precursors) are seen
floating nearby the pollen grains of Hn/?rcrrrre/isiirginimro
or filling up the reticulum. Ubisch-bodies (white arrows)
and other (granular) tapetal derivates will adhere to the
homogeneous film, while the exine cavities are Iack-
ing the sticky substances (Fig. 2 B). Some en-
tomophilous angiosperms with peculiar pollination
ecology (e.g. the so-called “Streukegel” in Ericrr)
are pollinated also by wind; thus they are “am-
phiphilous”, because their pollen grains are dry
rather than sticky. In such cases, as in Gnlorrtllirs
t i i u i l i s , the pollenkitt fills up preferentially the
cavities of the exine, while the pollen surface (e.g.
the tectum) remains without pollenkitt, and is rather
dry and not sticky (Fig. 1 B).
After leaving the pollen sac the mature pollen
grains of most entomophilous species are coated
with a thin film of pollenkitt; so they are able to
adhere as small clumps on hairs or on bristles of
flower-visiting insects or even on their smooth
chitinous parts (Hesse 1980 c).
The viscin threads are also of great importance in
the pollination of entomophilous angiosperms, but
do not act as a store for recognition substances in
the pollination process, and any allergenic function
pollen grain surface. TEhl x 17000. (B) Shortly before
anthesis the electron-dense pollenkitt totally covers the
tectum of Sogirtorin sngirrifolin and attaches the pollen
grain to the loculus wall (arrows). TEhl x 11 000.
seems to be impossible because of their chemistry.
It is very interesting indeed that they only occur in
some not closely related (mostly entomophilous,
but also ornithophilous, Fidrsiri) angiosperm
families. In most Onagraceae, in some Ericaceae,
but also in some (as known up to now) Caesal-
piniaceae (Skvarla et al. 1978, Cruden & Jensen
1979, Graham et al. 1980). Evidently the Onag-
raceae, and most probably also the other two
families, produce both pollenkitt and viscin threads
as vehicles for pollen cementing and pollen trans-
port (Fig. 3). Without doubt the viscin threads facili-
tate the pollen movement from the anther towards
the stigma. The origin of the viscin threads was
rather obscure until now. Evidently the threads are
neither sticky nor viscous like the pollenkitt, they
contain sporopollenin, and they are attached to the
exine (Skvarla et al. 1978, Hesse 1 9 8 0 ~ ) .The
threads occur first at the beginning of the formation
of the alveolar ektexine in Firclisin, long before the
tapetum starts to synthesize lipid globules as pol-
Grana 20
 Pollerikitt arid visciri threads 149
I_

* rj

=A-

Grnnn 20
150 h l . Hcssc
Fig. 5. The viscin threads on tetrads of Rhodode~idro~i
adhere like ropes to the bristles of a
sclrlipp~~iDtrchii
flower-vistinginsect. SEhI (the long bar indicates 10prn).
angiosperm pollen. The viscin threads are of the
same electron density as the exine and have the
same acetolysis resistance. The threads are partial-
ly coalesced with the ektexine. Most probably the
threads contain sporopollenin. Obviously the
threads are appendices of the ektexine and part of
the exine itself (Skvarla et al. 1978, Graham et al.
1980, Hesse 1980 6). Thus the term “exinal connec-
tions” coined by Cruden & Jensen (1979) is very
appropriate.
The size of the threads and their number are very
variable. In Onagraceae they are long, numerous,
thin and sculptured, while in Ericaceae they are
also long, numerous, thin, but smooth; in Caesal-
piniaceae there are short, thick and smooth threads.
Cruden & Jensen (1979) stated that the relative
number and length of the threads is related to pollen
clump size and this opinion seems to be more or less
adequate; but in my opinion the viscin threads are
not sticky, and so the “adhesion” of the threads on
the pollen surface results only from their coales-
Grana 20
cence with the ektexine. It is unknown, whether the
threads are coalesced with three or more pollen
grains or not.
The sculpture of the threads varies widely. Some-
times the threads are smooth throughout, as in most
Ericaceae and Caesalpiniaceae, while in Onag-
raceae there are knobs, furrows etc. The three-di-
mensional arrangement of the threads is very simi-
lar in Onagraceae and Ericaceae (Hesse 19806),
while there are fewer threads in the Caesal-
piniaceae. Despite the differences in morphology all
threads are undoubtedly strictly homologous. The
external morphology of the threads never depends
on the preparation method used, as there is no
modification of their fine structure or sculpture
after any. fixation or dehydration method, or even
acetolysis. If they are really made up of exinous
material, this would be expected. It has been re-
ported that pollen of some other angiosperm fa-
milies also have viscin threads, but this seems to be
a misinterpretation: The often cited “fibrils” of the
 Pollerikitt mid riscirz thrcritls 15 1

Table I. Origin arid characteristic prciiliarities of pollerikitt trritl visciii threcirls

Clrorncters Polleiikitt Virciri rlrrenils

Occurrence in In all families investigated Only in a few distinctly

angiosperms up to now entomophilous families
Origin Synthesis by plastids of the anther Like the material of the ektexine,
tapetum, mostly during the tetrad long before the pollenkitt synthesis
stage and later on
Chemistry A complex mixture of lipid substances Similar or equal to the ektexine,
containing sporopollenin

State of aggregation Fluid, viscous, sticky, partially with Firm, elastic, not fluid, not sticky,
crystalline inclusions without crystalline inclusions
Sculpture No sculpturing Often smooth surfaces, but partially
species-specific knobs or furrows
Fine structure Opaque, but with varying homogeneity The same homogeneity and electron-
and electron-density, partially density as the ektexine, without
with lamellae lamellae
Distribution on the In entomophilous angiosperms mostly The threads often originate near to
pollen surface as an electron dense, homogeneous film the apertures, but otherwise they
on the pollen surface; in anemophilous are not connected with the exine
angiosperms usually as small and
heterogeneous lumps in the cavities
of the exine
hlode of pollen The pollen grains (or tetrads) adhere The pollen grains (or tetrads) become
attachment only by their stickiness entangled by the threads, pollen
attachment by friction or adhesion

pollen grains of Strrlitziri (hlusaceae) are not viscin
threads or “exinal connections” at all, but evident-
ly filamentous cells between the pollen grains with-
out any connection with the exine (Skvarla et al.
1978, Hesse, in preparation).
The viscin threads fasten the pollen grains (or the
tetrads) like ropes, and the grains or tetrads become
entangled with their neighbouring grains or tetrads,
with insect hairs or bristles or even with the small
scales of the smooth surfaces of the flower-visiting
insects. Thus the pollen grains or tetrads adhere
only by friction (Fig. 5); although nectar vomited by
honey-bees sometimes reinforces the pollen
attachment (Hesse 1 9 8 0 ~ ) .It is obvious that all
viscin threads described up to now are products of
evolutionary specialization in the Onagraceae,
Ericaceae and Caesalpiniaceae. Perhaps long ago
the threads; like some other unusual features in
exine structure,. arose as a chance mutation. But
this structural change of the exine was not dis-
advantageous but rather a valuable adaptation for
biotic pollination; the modification of exine was
evidently successful during the evolution of the
entomophilous families Onagraceae, Ericaceae and
Caesalpiniaceae.
CONCLUSIONS
Pollenkitt and viscin threads differ widely not only
as to origin, structure, sculpture, chemistry, dis-
tribution on the pollen surface, occurrence in an-
giosperms, and also as to the mode of pollen
attachment (Table I). Because of these dis-
similarities and their simultaneous occurrence in
some families it is evident that the pollen attaching
vehicles are by no means identical; they are inde-
pendently derived. The formerly collectively used
concept “pollen cementing substances” can better
be regarded as an ecological, but by no means a
morphological term.
ACKNOWLEDGEhlENTS
This study was supported by the “Fonds zur Forderung
der wissenschaftlichen Forschung (projects 2441 and
3681)” and the “Kulturamt der Stadt Wen”.

Grana 20
152 M . Hesse
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Grana 20
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